Physicochemical, Microbiological, and Toxicological Characterization of Pâté Prepared from the Meat and Liver of Bullfrog (Aquarana catesbeiana) Carcasses.

The development of balanced, healthy, ready-to-consume, and easy-to-prepare products has led to the development of new food technologies. Despite their high commercial value, bullfrog (Aquarana catesbeiana) carcasses result in low yields, with the thighs being the most marketed in comparison to other carcass portions. In this sense, liver pâté is a traditional food consumed worldwide, mainly in European countries, and may be prepared by incorporating bullfrog meat by-products and certain viscera. In this context, the aim of the present study was to develop a pâté product based on a mixture comprising 50% grounded bullfrog torso meat and 50% liver paste, with each treatment incorporating 10% liver paste increments, totaling five final mixtures. The nutritional compositions and physicochemical, microbiological, and toxicological characteristics of each mixture were assessed. The dry matter percentage of the prepared product was determined to be 27.00%, while mineral content was 1.45%, lipid content was 4.00%, and total protein content was 20.00%. Finally, microbiological counts were in agreement with current food safety regulations. The developed pâté serves as a standard, recycling underused industrial materials, adding value to the production chain at low operational costs, creating a more accessible market, and promoting the popularization of this type of meat.

Fatty Acid and Multi-Isotopic Analysis (C, H, N, O) as a Tool to Differentiate and Valorise the Djebel Lamb from the Mountainous Region of Tunisia.

The objective of this study was to distinguish between the Tunisian Djebel lamb meat and meat from typical Tunisian production systems (PSs) through the fatty acids (FAs) profile and the stable isotope ratio analysis (SIRA). Thirty-five lambs from three different regions and PSs (D = Djebel, B = Bou-Rebiaa, and O = Ouesslatia) were considered for this purpose. The results demonstrated that the PS and the geographic origin strongly influenced the FA profile of lamb meat. It was possible to discriminate between the Djebel lamb meat and the rest of the dataset thanks to the quantification of the conjugated linoleic acids (CLA) and the branched chain FAs. Moreover, statistically different concentrations of saturated, monounsaturated and polyunsaturated FAs and a different n-6/n-3 ratio were found for grazing (D and BR) and indoor (O) lambs, making it possible to discriminate between them. As for the stable isotope ratio analysis, all parameters made it possible to distinguish among the three groups, primarily on the basis of the dietary regimen (δ(13C) and δ(15N)) and breeding area (δ(18O) and δ(2H)).

Alkaloid profiling of Italian alpine herbs using high resolution mass spectrometry (Orbitrap-MS).

Alpine pastures have not yet been extensively studied with regard to the presence of alkaloids in herbaceous plants. In this work the alkaloid profiles were characterised from a selection of 62 herbs collected from alpine pastures in north-eastern Italy. High-performance liquid chromatography coupled to a hybrid quadrupole-orbitrap mass spectrometer was used to evaluate the presence of 41 different alkaloids and quantify them using a targeted approach. Provisionally, 118 alkaloids were identified, including both free and glycosylated forms, making use of a homemade database and a suspect screening approach. rucifoline, gramine, heliotrine, lycopsamine, seneciphylline, and veratramine were quantified with concentrations ranging from 6 to about 100 µg kg-1 in 6 plants. Herbaceous plants belonging to the most well-represented plant families (Poaceae, 9 species; Asteraceae, 7; Lamiaceae, 6) showed distinct and characteristic alkaloid profiles and were correctly reclassified with an average accuracy of 85% (Partial Least Squares - Discriminant Analysis).

Detection of Antimicrobial Resistance, Pathogenicity, and Virulence Potentials of Non-Typhoidal Salmonella Isolates at the Yaounde Abattoir Using Whole-Genome Sequencing Technique.

One of the crucial public health problems today is the emerging and re-emerging of multidrug-resistant (MDR) bacteria coupled with a decline in the development of new antimicrobials. Non-typhoidal Salmonella (NTS) is classified among the MDR pathogens of international concern. To predict their MDR potentials, 23 assembled genomes of NTS from live cattle (n = 1), beef carcass (n = 19), butchers' hands (n = 1) and beef processing environments (n = 2) isolated from 830 wet swabs at the Yaounde abattoir between December 2014 and November 2015 were explored using whole-genome sequencing. Phenotypically, while 22% (n = 5) of Salmonella isolates were streptomycin-resistant, 13% (n = 3) were MDR. Genotypically, all the Salmonella isolates possessed high MDR potentials against several classes of antibiotics including critically important drugs (carbapenems, third-generation cephalosporin and fluoroquinolone). Moreover, >31% of NTS exhibited resistance potentials to polymyxin, considered as the last resort drug. Additionally, ≤80% of isolates harbored "silent resistant genes" as a potential reservoir of drug resistance. Our isolates showed a high degree of pathogenicity and possessed key virulence factors to establish infection even in humans. Whole-genome sequencing unveiled both broader antimicrobial resistance (AMR) profiles and inference of pathogen characteristics. This study calls for the prudent use of antibiotics and constant monitoring of AMR of NTS.

Hempseed By-Product in Diets of Italian Simmental Cull Dairy Cows and Its Effects on Animal Performance and Meat Quality.

Cull dairy cows are important contributors to total beef production in the USA and in Europe. Hempseed cake is a by-product of oil production and it is rich in unsaturated fatty acids (FA). This study aimed to investigate the effect of adding hempseed cake to the diet of Italian Simmental (IS) cull dairy cows on performances and meat quality. Twenty-six cull dairy cows were divided into three dietary groups: hay-based, corn silage-based and pasture-based diets. Within each group, the animals were equally divided into two treatments according to the protein source of the concentrate: hempseed cake (HEMP) or soybeans meal (SB). The trial lasted four months. HEMP showed similar in vivo performance and carcass characteristics, such as average daily gain (p > 0.05) and dressing percentage (p > 0.05), compared with SB. Meat characteristics, such as ether extract content and Warner−Bratzler shear force, were also similar between experimental groups (p > 0.05). Considering FA composition, HEMP showed similar saturated FA and polyunsaturated FA content (p > 0.05) but lower desirable fatty acids (p < 0.05) content and a tendentially lower hypocholesterolemic/hypercholesterolemic ratio (p < 0.10) than SFA. Hempseed cake can substitute soybean in the diet of cull dairy cows without effects on performance or meat quality.

Gas Chromatography Combustion Isotope Ratio Mass Spectrometry to Detect Differences in Four Compartments of Simmental Cows Fed on C3 and C4 Diets.

Fatty acids (FAs) metabolism in animals represents an important field of study since they influence the quality and the properties of the meat. The aim of this study is to assess the possibility to discriminate the diets of cows in different animal compartments and to study the fate of dietary FAs in the bovine organism, using carbon isotopic ratios. Five FAs, both essential (linoleic and linolenic) and non-essential (palmitic, stearic, and oleic) in four compartments (feed, rumen, liver, meat) of animals fed two different diets (based on either C3 or C4 plants) were considered. For all compartments, the carbon isotopic ratio (δ13C) of all FAs (with few exceptions) resulted significantly lower in cows fed on C3 than C4 plants, figuring as a powerful tool to discriminate between different diets. Moreover, chemical reactions taking place in each animal compartment result in fraction processes affecting the δ13C values. The δ13CFAs tendentially increase from feed to meat in group C3. On the other hand, the δ13CFAs generally increase from rumen to liver in group C4, while δ13CFAs of rumen and meat are mostly not statistically different. Different trends in the δ13CFAs of the two groups suggested different FAs fates depending on the diet.

The Influence of the Type of Dry-Cured Italian PDO Ham on Cathepsin B Activity Trend during Processing.

Cathepsin B activity was measured during processing in hams originating from the main Italian prosciutto PDOs: Parma, San Daniele and Toscano. Sixty-five heavy pig thighs, from sixty-five Italian large white x Italian Landrace pigs bred and slaughtered in the same conditions were considered. Five thighs represented the post-mortem control time. The other 60 were distributed one plant per PDO, following a balanced plan. The thighs were sampled at the biceps femoris in groups of four per plant in the following ripening phases: salting, resting, drying, greasing, end of curing. The activity of the Cathepsin B (U/g protein) was determined by means of fluorescence measurements. The Cathepsin B ripening trend of the various PDOs was significantly different, particularly during the initial and mid-curing stage. This activity correlates with the proteolysis index through a PDO dependent pattern, indicating that different processing conditions can influence the quality of prosciutto, since they determine its biochemical development.

Using Bioelements Isotope Ratios and Fatty Acid Composition to Deduce Beef Origin and Zebu Feeding Regime in Cameroon.

The purpose of this study was to address the lack of knowledge regarding the stable isotopic composition of beef from zebu cattle reared in tropical Africa. Sixty beef carcasses belonging to the most common zebu breeds (Goudali, white Fulani, and red Mbororo) were selected and classified according to their subcutaneous fat color (white, cream or yellow). The stable isotope ratios of five bioelements-H, O, C, N, and S-in muscle fractions and the fatty acids composition were analyzed. Zebu meat from Cameroon shows peculiar δ13C values, related to the almost exclusive intake of grazed tropical grasses with photosynthetic cycle C4. It also shows δ2H and δ18O values higher than those reported in other areas of the world and correlated with the isotopic composition of animal drinking water. The white subcutaneous fat ("white type") zebu showed higher δ2H and lower δ13C than the "yellow type", that is correlated with a higher content of polyunsaturated fatty acid (PUFA) and a lower amount of saturated fatty acid (SFA) and monounsaturated fatty acid (MUFA). Multielement analysis seems to provide promising results for tracing the regional origin of Cameroon beef and some aspects of the livestock system, such as the nutritional status of the animals.

Fatty acids stable carbon isotope fractionation in the bovine organism. A compound-specific isotope analysis through gas chromatography combustion isotope ratio mass spectrometry.

The contribution of dietary fatty acids to the quality of the meat and their path through the bovine organism is currently the subject of a lot of research. Stable isotope ratio analysis represents a powerful tool for this aim, one that has not been studied in depth yet. In this work, for the first time, the carbon isotopic ratios of six fatty acids (myristic 14:0, palmitic 16:0, stearic 18:0, oleic 18:1n-9, linoleic 18:2n-6 and linolenic 18:3n-3 acids) in different matrixes (diet, rumen, duodenal content, liver and loin) were analysed through gas chromatography combustion isotope ratio mass spectrometry. Moreover, the quantification of the single fatty acids was carried out, providing important information supporting the carbon isotopic ratio results. The variation in the concentration of the fatty acids in the different matrices depends on the chemical modifications they undergo in the sequential steps of the metabolic path. GC-C-IRMS turned out to be a powerful tool to investigate the fate of dietary fatty acids, providing information about the processes they undergo inside the bovine organism.

Influence of dietary inclusion of tannin extracts from mimosa, chestnut and tara on volatile compounds and flavour in lamb meat.

Tannins are compounds able to form complexes with proteins limiting their ruminal degradation and thus the synthesis of some odour-active compounds may be inhibited. Tannins are broadly divided in condensed tannins (CT) and hydrolysable tannins (HT). The study aimed to assess the influence of dietary inclusion of three commercial tannin extracts, namely mimosa (Acacia mearnsii; CT), chestnut (Castanea sativa; HT) or tara (Caesalpinia spinosa; HT) on volatile profile and flavour of meat and kidney fat from lambs. Comisana male lambs were divided into four groups (n = 9 each) and fed for 75 days with a concentrate-based diet (CON) or CON supplemented with 4% of one of the tannin extracts. Tannins reduced "pastoral" odour in perirenal fat of lambs the meat of which was characterized by a very low perception of this attribute. It may be assumed that p-cresol and 8-methylnonanoic acid mostly contributed to "pastoral" odour expression in the diet without condensed or hydrolysable tannins.

Pungency perception and liking for pasta filata cheeses in consumers from different Italian regions.

We aimed to investigate whether the sensitivity to capsaicin had an effect on pungency perception of pasta filata cheeses. In addition, we assessed the effect of several individual variables (gender, age, consumer provenance, PROP status and personality traits) on both consumer liking and choice for pasta filata cheeses. Four pasta filata cheeses at two ripening times and 272 subjects from three different Italian locations were used. Based on sensory data, the PCA discriminated sweet (ripened at 2 months) from pungent cheeses (ripened at 12 months). Cheese pungency perception increased with increasing sensitivity to capsaicin (P < 0.001). There was no clear relationship between sensitivity to capsaicin or sensitivity to cheese pungency with cheese liking, whereas a number of other aspects, including sensory attributes and individual consumer characteristics, such as consumer provenance (P < 0.001), gender (P < 0.001) and food neophobia (P < 0.05), affected the liking for different pasta filata cheeses. Consumers who were younger (18-30 y.o.; P < 0.05), female (P < 0.001), unfamiliar with pungent foods (P < 0.0001) and consumers from Potenza (P = 0.001) more often declared to prefer sweet over pungent cheese, whereas in terms of real choice supertasters and neophobic subjects chose the sweet option more often (P = 0.01). In conclusion, sensitivity to capsaicin affected pungency perception in pasta filata cheese, whereas no clear relationship was observed between pungency perception and liking. The contribution of cheese and consumer characteristics on cheese liking and choice in addition to pungency, was confirmed by the differences in declared and real choice for sweet or pungent cheeses.

Effect of Heat Stress on Dairy Cow Performance and on Expression of Protein Metabolism Genes in Mammary Cells.

The aim of this study was to assess the effect of heat stress on dairy cow performance and on the expression of selected genes involved in milk protein metabolism. Eight Italian Holstein Friesian cows were kept under thermoneutral conditions (temperature-humidity index (THI) < 72, CON) for 8 days and under mild heat stress conditions (72 < THI < 78, HS) for an additional 8 days. The rectal temperature, feed intake, and milk yield were recorded during the last 3 days of the CON and HS periods. During the same time period, milk samples were collected to assess the composition and expression of selected genes involved in milk protein metabolism. Gene expression analyses were performed on somatic cells from milk, which are representative of mammary tissue. In terms of dairy cow performance, HS resulted in lower milk and protein yields and feed intake but higher rectal temperature than for CON (p < 0.05). Under HS, there were greater abundances of HSPA1A (p < 0.05) and BCL2 (p < 0.05), compared to CON, but similar levels of CSN2 (p > 0.05), CSN3 (p > 0.05), HSPA8 (p > 0.05), and STAT5B (p > 0.05) mRNA. Mild heat stress reduced the performance of dairy cows without affecting the expression of genes coding for caseins.

Environmental Sustainability Assessment of Dairy Farms Rearing the Italian Simmental Dual-Purpose Breed.

This study aimed to assess the environmental footprint of dairy farms rearing a dual-purpose breed, and to evaluate, through alternative scenario analyses, the fattening of calves and the cultivation of hemp as strategies for reducing the environmental impact of these farms. Eleven farms were evaluated for global warming (GWP), acidification (AC) and eutrophication (EUP) potential. The Life Cycle Assessment method with three scenarios, REAL, based on real data, BEEF, where calves were fattened in farm, and HEMP, where hemp was cultivated in farms, were considered. If referred to 1 m2 of utilizable agricultural land, the GWP, AC and EUP were 1.6 kgCO2eq, 21.7 gSO2eq and 7.1 gPO43-eq, respectively. If referring to 1 kg of fat and protein corrected milk, the emissions were 1.1-1.4 kgCO2eq, 14.8-19.0 gSO2eq, and 5.0-6.4 gPO43-eq, depending on the allocation method adopted. The emissions were associated positively with culling rate and negatively with production intensity. In BEEF and HEMP scenarios, the emissions were reduced by 8-11% and by 1-5%, respectively. Fattening the calves, evaluating the cultivation of alternative plants and improving the productive and reproductive efficiency of animals could be effective strategies for reducing the environmental footprint of the farm.

Influence of dietary cardoon meal on volatile compounds and flavour in lamb meat.

Cardoon meal is a by-product retained after oil extraction from the seeds of cultivated Cynara cardunculus var. altilis that has been proposed as a valuable resource for animal feeding. The study aimed to assess the influence of its dietary inclusion on volatile profile and flavour of meat and kidney fat from lambs. Fifteen Sarda × Comisana male lambs were randomly divided in two groups and fed for 75 days with a concentrate-based diet containing 15% cardoon meal (CMD, n = 7) or dehydrated alfalfa (CON, n = 8). Cardoon meal inclusion reduced the "barnyard/animal" odour perceived by a trained panel in kidney fat, which could be associated with the aromatic compound p-cresol (4-methylphenol), detected only in CON diet. Considering the other aroma volatiles regarded as the main contributors for the characteristic lamb flavour, both diets were characterized by moderate to low levels of 4-methyloctanoic acid, skatole and indole while 4-methylnonanoic acid and 4-ethyloctanoic acid were not detected.

Do Dairy Farming Systems Differ in Antimicrobial Use?

The quantitative assessment of antimicrobial use (AMU) in food-producing animals contributes to the provision of essential information for developing relevant and effective policies to reduce use and to control antimicrobial resistance. Information on AMU is available mainly for intensive dairy farming systems and specialized high-yielding breeds. The aim of this study is to investigate AMU in different dairy farming systems by comparing the treatment incidence in mountain farms with specialized high-yield dairy breeds or with dual-purpose breeds raised for milk production to the treatment incidence in lowland farms with specialized high-yield dairy breeds or with dual-purpose breeds raised for milk production. Significant differences were found only between the overall treatment incidence, as well as the treatment incidence of highest-priority critically important antimicrobials for human medicine, in lowland farms with high-yielding breeds and mountain farms with dual-purpose breeds. Mountain farms have a generally lower milk production and smaller herd size than lowland farms, provide cows with access to pasture, and limit concentrates in the diet. These management practices and the use of local/dual-purpose breeds could reduce the risk of production diseases and the consequent need for AMU.

Fatty Acid Profiles of Cow's Milk and Cheese as Affected by Mountain Pasture Type and Concentrate Supplementation.

The aim of this trial was to assess the effect of pasture type and concentrate supplementation on the fatty acids (FA) composition of milk and cheese obtained during summer grazing on mountain pasture. Seventy-two Italian Simmental dairy cows were assigned to two groups that differed by the amount of concentrate supplementation: 3.0 kg/head/d (HIGH) vs. 1.5 kg/head/d (LOW). The dairy cows grazed on a Poion alpinae alliance pasture (PAST1), and subsequently they grazed on a Seslerion caeruleae alliance pasture (PAST2) for 10 d each. In the last three days of each experimental period, milk samples were collected immediately before each cheese-making event. Cheese samples were collected from each cheese loaf after 60 d of ripening. LOW showed higher isoFA, FA intermediates of the ruminal biohydrogenation, C18:3 c9,c12,c15, and total polyunsaturated fatty acid (PUFA) levels than HIGH. The pasture type had a more limited effect on FA composition of milk than concentrate level and was mainly related to monounsaturated fatty acids (MUFA), which were higher in PAST1 than PAST2 (p < 0.05). In cheeses, these differences were reduced. The phytanic acid and phytanic isomer ratio (SRR/RRR) in milk were not affected either by supplement level (p > 0.05) or by type of pasture (p > 0.05). Increasing the concentrate offered to dairy cows from 1.5 to 3.0 kg/d did not markedly influence the level of PUFA in cheeses produced during summer grazing on high mountain pasture.

Isotopic and elemental profiles of Mediterranean buffalo milk and cheese and authentication of Mozzarella di Bufala Campana PDO: An initial exploratory study.

Mozzarella di Bufala Campana (MBC) is a PDO cheese produced from whole buffalo milk in specific regions of southern Italy. Due to the high price and the limited amount of buffalo milk, MBC is potentially subject to mislabelling. Stable isotope ratio analysis combined with elemental analysis is one powerful technique for detecting the authenticity of PDO cheeses. Here, the elemental and isotopic profiles of authentic samples of buffalo milk and the corresponding MBC samples collected in the reference area in winter and summer are presented in an initial exploratory study. By merging MBC-PDO samples with non-PDO samples of buffalo mozzarella produced both inside and outside the reference area, a model was developed to classify product categories for this cheese. Despite the differences occurring during processing, along with differences in the season and production area, the model was effective in distinguishing PDO and non-PDO mozzarella, particularly when non-PDO cheeses were made outside the MBC reference area.

Metagenomic profiles of different types of Italian high-moisture Mozzarella cheese.

The microbiota of different types of Italian high-moisture Mozzarella cheese produced using cow or buffalo milk, acidified with natural or selected cultures, and sampled at the dairy or at the mass market, was evaluated using a Next Generation Sequencing approach, in order to identify possible drivers of the bacterial diversity. Cow Mozzarella and buffalo Mozzarella acidified with commercial cultures were dominated by Streptococcus thermophilus, while buffalo samples acidified with natural whey cultures showed similar prevalence of L. delbrueckii subsp. bulgaricus, L. helveticus and S. thermophilus. Moreover, several species of non-starter lactic acid bacteria were frequently detected. The diversity in cow Mozzarella microbiota was much higher than that of water buffalo samples. Cluster analysis clearly separated cow's cheeses from buffalo's ones, the former having a higher prevalence of psychrophilic taxa, and the latter of Lactobacillus and Streptococcus. A higher prevalence of psychrophilic species and potential spoilers was observed in samples collected at the mass retail, suggesting that longer exposures to cooling temperatures and longer production-to-consumption times could significantly affect microbiota diversity. Our results could help in detecting some kind of thermal abuse during the production or storage of mozzarella cheese.

Stable isotope ratios of H, C, O, N and S for the geographical traceability of Italian rainbow trout (Oncorhynchus mykiss).

Multielement isotope ratio was assessed in fat and defatted (protein) fillet of 130 rainbow trout, reared with feed incorporating a high or low fish content in 20 Italian farms, focusing on two northern Italian regions (Friuli Venezia Giulia and Trentino). The aim was to investigate the ability of isotopic analysis to trace the geographical origin of trout, also according to the type of feed. The carbon, nitrogen and sulphur isotope ratios of feed and fillet were highly positively correlated both within each matrix (feed or fillet) and between the two matrices and negatively correlated with the δ2H and δ18O of feed and the δ2Hfat. The δ2Hprotein and δ18Oprotein were positively interrelated with the δ18O of tank water. By applying the tested Partial Least Squares - Discriminant Analysis multiclass model (85 fillets) to the validation dataset (45 fillets), 91% accuracy was obtained for the two Italian regions.

Concentrate Supplement Modifies the Feeding Behavior of Simmental Cows Grazing in Two High Mountain Pastures.

During grazing on Alpine pastures, the use of concentrates in dairy cows' diet leads to a reduction of the environmental sustainability of farms, and influences the selective pressure on some plant species. In order to minimize the use of concentrates, it is imperative to obtain data on the grazing behavior of cows. The aim of this study was to assess the effect of concentrate levels on the behavior of dairy cows during grazing. One hundred and ten lactating Italian Simmental cows, that sequentially grazed two pastures characterized by Poion alpinae (Poion) and Seslerion caeruleae (Seslerion) alliance, were considered. For each pasture, eight cows were selected and assigned to two groups: High and Low, supplemented with 4 kg/head/d, and 1 kg/head/d of concentrate respectively. Cows were equipped with a noseband pressure sensor and a pedometer (RumiWatch system, ITIN-HOCH GmbH) to assess grazing, ruminating, and walking behavior. In addition, the plant selection of the animals was assessed. On Poion, increased supplement intake caused a more intense selection of legumes, without affecting feeding and walking times. On Seslerion, grazing time was higher in Low than High. Grazing management in alpine region must take into account the great variability of pastures that largely differ from a floristic and nutritional point of view.